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Effect of Mexican Oregano ( Lippia Graveolens Kunth)

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Effect of Mexican Oregano ( Lippia Graveolens Kunth) Annual Research & Review in Biology 4(23): 3470-3491, 2014 SCIENCEDOMAIN international www.sciencedomain.org Effect of Mexican Oregano (Lippia graveolens Kunth) on Streptozotocin Induced Diabetic Mice and its Role in Regulating Carbohydrate Metabolic Enzymes and Their inhibitory Effect on the Formation of Advanced Glycation end Products Rosa Martha Perez Gutierrez1* 1Research Laboratory of Natural Products, School of Chemical Engineering and Extractive Industries-IPN, Unidad Profesional Adolfo Lopez Mateos, Zacatenco, D. F. CP 07758, Mexico. Author’s contribution This whole work was carried out by author RMPG. Received 29th April 2014 rd Original Research Article Accepted 23 May 2014 Published 19th June 2014 ABSTRACT Aim of the Study: The aim was to evaluate the effect of extracts of the leaves of Lippia graveolens in streptozotocin-induced mildly diabetic (MD) and severely diabetic (SD) miceand on the formation of advanced glycation end products (AGEs). Study Design: Diabetes has emerged as a major global challenge in healthcare delivery, particularly in recent times, with the global incidence reaching epidemic proportions. Because of this there is an increasing demand to research for natural products with antidiabetic activity. Place and Duration of Study: Laboratory of Natural Products Research. School of chemical engineering, National Polytechnic Institute between August 2013 and April 2014. Methodology: Extracts were orally administered to MD and SD rats for 30 days, and a set of biochemical parameters were studied: glucose, cholesterol, triglycerides, lipid peroxidation, glycogen, SOD, CAT, GSH, GPX, glucose-6-phosphatase, glucokinase, hexokinase activities, SGOT, SGPT, ALP and insulin level. In this study, we investigate the ____________________________________________________________________________________________ *Corresponding author: Email: [email protected]; Annual Research & Review in Biology, 4(23): 3470-3491, 2014 inhibitory effects In vitro on the formation of specific AGE representatives including AGEs- BSA formation, Amadorin activity, methylglyoxal (MGO). In vivo was to investigate the effect of extracts on oxidative stress, glycation of hemoglobin and MGO, glycolaldehyde (GA) levels, TBA-reactive substance level in kidney mitochondrial instreptozotocin-induced diabetic mice. Results: Methanol extract (LG-M) reduced the intake of food, water and body weight loss as well as levels of blood glucose, serum cholesterol, triglyceride, lipoprotein and increase HDL-cholesterol, antioxidant enzymes, improves TBARS–reactive substance, marker enzymes of hepatic function. These results, support that improves glucose metabolism by reducing insulin resistance. LG-M is an inhibitor of fluorescent AGE, methylglyoxal and the glycation of haemoglobin. Conclusions: This study demonstrated that Lippia graveolens possesses considerable anti-AGE and hepatoprotective role, inhibits hyperglycemic, hyperlipidemic and oxidative stress indicating that these effects may be mediated by interacting with multiple target operating in diabetes mellitus. Keywords: Antidiabetic; hyperlipidemia; glycation; lippie graveolens; antiglycation. 1. INTRODUCTION Diabetes is a serious metabolic disorder with micro and macro vascular complications resulting in significant morbidity and mortality. Diabetes mellitus is principally characterized by insulin resistance (IR) and elevated blood glucose levels [1]. Prolonged hyperglycemia contributes importantly to the pathogenesis of diabetic complications by increasing protein glycation, leading to the gradual buildup of advanced glycation end products (AGEs) in body tissue [2]. The complex, fluorescent AGE molecules formed during Maillard reaction can lead to protein cross-linking, which contributes to the development and progression of various diabetic complications [3]. Diabetes is characterized by abnormal lipid and protein metabolism, along with specific long- term complications affecting the retina, the kidney and the nervous system mainly [4]. Oxidative stress and advanced glycation endproducts (AGEs) formation induced by hyperglycemia are known to influence diabetic renal changes and nephropathy [5]. The number of people with diabetes is increasing worldwide due to population growth, aging, urbanization, increasing prevalence of obesity, calorie rich diet and physical inactivity. Current treatments, although provide a good glycemic control do little preventing complications [6]. Besides, most of the prescribed hypoglycemic drugs or insulin are associated with unwanted side effects. Herbal medicines are an option because of their comparably therapeutic effects and nontoxic side effects [7]. Lippia graveolensis, commonly known as “Mexican oregano”, distributed widely in México, Central and South America. It is widely used in Mexico as food seasoning and a folk remedy.Aerial part ofthis species is used as an antiseptic, antipyretic, analgesic, abortive, antispasmodic, anti-inflammatory agent and for the treatment of menstrual disorders and diabetes [8]. L. graveolensis used in Central America for the treatment of gastrointestinal and respiratory diseases, and as antipyretic, digestive, anti-inflammatory and antifertility drug [9]. The essential oil presented antifungal activity which can be attributed to the presence of carvacrol, α-terpinyl acetate, cymene, thymol, pinene and linalool, which are already known to exhibit antimicrobial activity [10]. Most of the studies on the chemical composition of 3471 Annual Research & Review in Biology, 4(23): 3470-3491, 2014 L. graveolens have mainly focused on the terpene compounds contained in the essential oil as thujene, β-pinene, m-cymene, 1,8-cineole, α-terpinyl acetate, linalool, camphor, terpinen- 4-ol, bornyl acetate, carvacrol, thymol, cadina-4(5),10(14)-diene, guaia-1(10),11(12)-diene, isocaryophyllene, humulene, aristol-1(10)-ene, α-bisabolol, β-bisabolene, β-candinene, himachalene and cedrene [10]. In vitro studies have shown that essential oil from methanolic extract have antioxidant and antimutagenic properties [11]. L. graveolens has been reported that essential oils with a highcarvacrol content displayed antimicrobial [12] and antiviral effects [13]. An investigation of the leaves of L.graveolens from Guatemala provided iridoid and secoiridoid glucosidesas loganin, secologanin, secoxyloganin, dimethyl secologanoside, loganic acid, 8-epi-loganic acid, caryoptosidic acid, caryoptoside, lippiosideI and II [14]. In several phytochemical studies on this plant led to the isolation of thirty-oneflavonoids [15]. Furthermore, the aqueous extract of leaves inhibited some trophozoites [16]. In addition, flavanone(-)(2S)-5,6,7,3',5'-pentahydroxyf1avanone-7-O-β-D- glucopyranoside shown anti-inflammatory and cytotoxic activities [17]. Although oregano is popular around the world its beneficial and/or adverse effects on human health have not been scientifically determined yet. No previous study has, for instance, so far reported on the systematic investigation and evaluation of the antidiabetic and antiglycation activities of oregano. To the authors' knowledge, the present work is the first attempt to investigate the protective effects of oregano on diabetes and its complications on the functions of the liver, kidney, and pancreas. 2. MATERIALS AND METHODS 2.1 Plant Material Lippia graveolens Kunth belong to the Verbenaceae family, leave were collected in Morelos state and were taxonomically authenticated in the Herbarium of Escuela Nacional de Ciencias Biologicas, Instituto Politécnico Nacional. A voucher specimen of the plant is stored for reference (No. 9289). Leaves of L graveolens was air dried and the ground (2kg) was extractedtwice with hexane, chloroform and methanol each for 3h. The extracts were evaporated invacuum to generate a residue (28.5, 36.2, 23.4 g residue respectively). 2.2 Experimental Animals 2.2.1 Experimental animals The study was conducted in CD1 male mice, weighing about 25-30g. Before and during the experiment, animals were fed a standard laboratory diet (Mouse Chow 5015, Purina) with free access to water. The experiments reported in this study were carried following the guidelines stated in Principles of Laboratory Animal Care (National Institute of Health publication (NIH) 85-23, revised 1985 and the Mexican Official Normativity (NOM-062-Z00- 1999).All experiments and protocols described in present study were approved by the Institutional Animal Ethical Committee of Escuela Nacional de Ciencias Biologicas-IPN (Regd. No. 735). 2.3 Induction of Severe Diabetes Severe diabetes was induced in overnight fasted male mice by a single intraperitoneal injection of streptozotocin (Sigma Chemical Company, St. Louis, MO, USA), at a dose of 50 mg/kg body weight dissolved in cold citrate buffer (pH 4.5) [18]. Hyperglucemia was 3472 Annual Research & Review in Biology, 4(23): 3470-3491, 2014 confirmed by measuring glucose 72h after the streptozotocin shot and 7 days after injection, confirming a high glucose level. Rats with permanent high fasting blood glucose level >300mg/dl were included for the experiments. 2.4 Induction of Mildly Diabetes Mild diabetes was induced in overnight fasted mice by administering a single dose of freshly prepared solution of streptozotocin (STZ), 45mg/kg b.w. i.p) in 0.1mol/l cold citrate buffer (pH 4.5), 15 min after the intraperitoneal administration of 120mg/kg nicotinamide (Sigma Chemical Company, St. Louis, MO, USA). The STZ treated animals were allowed to drink 5% glucose solution over night to overcome
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